Preparation, characterization, and in vitro enzymatic degradation of chitosan-gelatine hydrogel scaffolds as potential biomaterials.

The crosslinking of chitosan (CHT) and gelatin (GEL) accomplished with N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS) was investigated and optimized in relation to hydrogels stability by varying the CHT/GEL mass ratio and the EDC/NHS molar ratio at different and constant EDC concentrations. Hydrogels were also fabricated in the presence of α-tocopherol to assess the release mechanism of a lipophilic drug from a highly-hydrophilic CHT/GEL hydrogel network. Alterations in the physico-chemical properties of hydrogels were characterized by differential scanning calorimetry (DSC) and fourier transform infrared spectroscopy (FTIR), and their biostability was studied within a simulated body-fluid solution (PBS of pH 7.4) at 37 °C for 24 h by evaluating the degree of swelling, followed by topography and morphology characterization using scanning electron microscopy (SEM). The analysis confirmed the formation of a modulated hydrogels porosity using different freezing temperatures prior to lyophilization. The in vitro degradation behaviors of the hydrogels were investigated for up to 5 weeks using collagenase, lysozyme, and N-acetyl-β-D-glucosaminidase by monitoring the weight-losses of hydrogels and their degradation products, being identified by UV-Vis spectroscopy and high-performance liquid chromatography (HPLC) as well as the pH monitoring of degraded solutions. It was observed that an inner morphological hydrogel structure influences their swelling and degradation behavior, which is additionally reduced by in-gel-embedded α-tocopherol because of hydrophobic interactions with their constituents, and hindering the effect on collagenase activity.